Evaluation of Anticoagulant and Antiplatelet Activity of Pisum sativum Pod Extract
The current study evaluates the anticoagulant and antiplatelet activities of Pisum sativum pod extract (PSPE). PSPE revealed similar protein banding pattern under both reducing and nonreducing conditions on SDS-PAGE, suggesting the presence of monomeric proteins in the extract. PSPE showed proteolytic activity as it hydrolyzed casein and gelatin and the specific activity was found to be 0.16 and 0.21 units/mg/min respectively. The proteolytic activity of PSPE was completely abolished by PMSF but not IAA, EDTA and 1,10, Phenanthroline suggesting the presence of serine protease. Further, PSPE was evaluated for anticoagulation efficiency. Interestingly, PSPE increased the clotting time of citrated human plasma from the control 220 sec to 460 sec suggesting its anticoagulant property. Furthermore, PSPE specifically prolonged the clotting time of APTT but not PT revealed the interference in intrinsic pathway of coagulation cascade. In addition, PSPE exhibited anti-aggregation activity by inhibiting agonists such as ADP and epinephrine induced platelet aggregation in a dose dependent manner and the detected aggregation inhibition was found to be 58% and 82% respectively. In addition, PSPE preferentially hydrolyzed Aα and Bβ chain of human fibrinogen and all the chains of fibrin clot. Fibrinogenolytic and fibrinolytic activity of PSPE was inhibited by PMSF but not IAA, EDTA and 1,10, Phenanthroline suggesting the presence of serine protease in the extract. Moreover, PSPE did not hydrolyze RBC cells signifying its non-toxic property.